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Qualitative and semi-quantitative comparisons of calcium content in the developing human mandible were performed by means of microradiographic and histophotometric analysis. Differences in calcium content between enamel, calcified cartilage, chondroid tissue and dentin are significant at the 1% level. Chondroid tissue and woven bone are almost similarly mineralized tissues.
To investigate the cell-matrix interrelation and the structure and permeability of the junctional complexes of secretory ameloblasts, molar tooth germs from kittens were examined by means of scanning electron microscopy, routine thin sections and freeze-fracture replication. Scanning electron microscopy showed remarkably dissolved growth fronts of enamel in materials that had been fixed with glutaraldehyde and then subjected to EDTA perfusion for 10 min. By the action of EDTA, intercrystallite spaces in rod and interrod enamel were prominently widened, and their longitudinal ends of crystallites displayed irregular and extremely sparse structures. In enamel rods surrounded entirely by interrod enamel, and in enamel rods of the typical key hole shape with successive interrod enamel participation, the most striking dissolution of crystallites occurred at the boundaries between rod and interrod enamel, where broad expanses of rod-sheath spaces were observed. In thin sections, the Tomes processes of secretory ameloblasts occupying the above rods were rectangular or variations of a rectangular shape, respectively; and interameloblast spaces opened to the enamel growth fronts, which corresponded to the junction between rod and interrod enamel. In enamel rods standing in regular rows and showing the typical arcade shape, the centers of the rods were drastically dissolved and exhibited single and deep slits, whereas the boundaries between rod and interrod enamel showed no wide furrows. The Tomes processes occupying such arcade-shaped rods were typically triangular, and the interameloblast space always joined the type-1 face of process, which is responsible for enamel rod formation. Secretory ameloblast possessed two sets of junctional complexes at the proximal and distal ends of the cell body. The distal one was situated proximally to the Tomes process. Freeze-fracture replication demonstrated the functional structures of these junctions: the proximal junction was fascia occludens, and the distal one incomplete zonula occludens with many free-ending tight junctional strands and interstrand spaces or a less developed irregular junction.
Young CD-1 mice, 4 days old, exposed to 0.1% nicotine sulfate on gestational days 6-20 were compared with untreated pups of the same age to determine its effect on the development of mandibular first molars. Pregnant mice were given intraperitoneal injections of nicotine at a dose of 1.67 mg/kg/day. Pups were then decapitated, their entire mandibles were excised, routinely prepared and embedded in paraffin, sectioned in the frontal plane and stained with hematoxylin and eosin for histological examination of developing lower first molars. The results demonstrated that the process of odontogenesis appears retarded in nicotine-treated animals while the molars of the control group revealed dentin and enamel formation. It was concluded that nicotine has a detrimental effect on molar development. Nicotine may interfere with cellular maturation of the tooth germ indicating that this effect is prenatal and extends postnatally.
The earliest stages of development and differentiation of deciduous and permanent teeth were studied in prenatal and perinatal tarsiers. Both deciduous and successional teeth develop at loci for I1, I2, C, P2-4. Relationships among the dental lamina, enamel organs, and successional lamina were used to determine tooth homologies. During the second half of prenatal life, dP2 and dI2 undergo abnormal development. Their dentin becomes fragmented, and these teeth are shed or resorbed without eruption during the perinatal period. Our study provides evidence of an intermediate phase in evolutionary tooth loss. These findings do not corroborate recent hypotheses of primate phylogeny based mainly on tooth eruption patterns.
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Fetuses of known age, collected from 20 days' gestation to term, were used to characterize the chronology of deciduous tooth development within the right mandible of swine. Tooth development was first observed at 32 days' embryonic development, with the differentiation of the deciduous third molar. Bud stages for the remaining deciduous teeth differentiated within the period of 32 to 38 days of embryonic development. Although the initial histological appearance of these teeth covered a short period of time, it was apparent that each tooth continued to develop at its own rate. The deciduous second incisor and first molar reached a stage of enamel formation by the 80th and 86th day of fetal development. This is a much later stage than previously recorded for beginning enamel formation. The stages of tooth development and enamel formation for each tooth are summarized. A previous report on the distribution of the dental lamina and deciduous tooth development in the mandible of the domestic pig combined with the information presented in this report on tooth chronology provide much of the information required for future studies using the domestic pig in dental research. A fetus observed at the 74th day of development demonstrated a tooth bud for the deciduous first premolar. The development of this tooth was followed closely throughout the remainder of fetal development with the cap stage representing its most definitive form at 110 days' development. The suggested deciduous origin for this tooth could result in a reevaluation of the nomenclature for the dental formula of swine.
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Partially mineralized enamel matrix removed from bovine incisor tooth germs contains proteolytic activity capable of completely degrading enamel matrix proteins at neutral and slightly alkaline pH. The protease activity also degrades common protease substrates such as casein and azocoll with optimal activity at pH 8-9. Inhibitor studies revealed that the enzyme involved is a serine protease of the chymotrypsin family.
Eight groups of dental tissues were mechanically dissected from the mandibles of one-year-old steers; they were then defatted and decalcified in HCl. The noncollagenous proteins were extracted with various solvents from collections of tissue and bio-assayed for osteo-inductive activity. Collectively, the hard tissue (dentin, enamel, and cementum) noncollagenous proteins were fractionated by molecular sieve chromatography, hydroxyapatite affinity chromatography, and ion exchange chromatography. Osteo-inductive activity of each protein fraction was determined by implantation in the quadriceps muscle pouch of mice. The quantity of bone was measured by computerized image analysis. From 71% to 83% of 41 implants of dental hard tissues induced bone formation. The quantity of bone was greater from unerupted than from erupted teeth. Dental soft tissues that had no osteo-inductive activity were rich in a 14-kDa protein, presumably matrix gamma-carboxyglutamic acid-rich proteins. Proteins with Mr of from 15 to 28 kDa were associated with osteo-inductive activity. Components with Mr greater than 28 kDa had no activity. These observations suggest that bovine teeth have a selection of osteo-inductive proteins that is comparable in range of MW to bovine bone morphogenetic protein.
The periodontal ligament (PDL) is a unique tissue that is crucial for tooth function. However, little is known of the molecular mechanisms controlling PDL function. To characterize PDL cells at the molecular level, we constructed a cDNA library from bovine PDL tissue. We then focused on the isolation of S100 calcium-binding proteins (CaBPs), because they mediate Ca2+ signaling and control important cellular processes such as differentiation and metabolism. We screened the PDL cDNA library with a mouse S100A4 cDNA, and cloned the bovine cDNAs of two S100 CaBPs (S100A4 and S100A2). In northern blotting analysis, the highest expression of S100A4 was detected in PDL from erupted teeth (PDLE). PDL from teeth under eruption (PDLU) showed a lower expression of S100A4, and its expression in gingiva was faintly detectable. S100A4 expression was also high in the pulp tissue followed by the dental papilla of the tooth germ. S100A2 expression was high in PDLE and gingiva. Interestingly, only PDLE exhibited a high expression of both S100A4 and S100A2. PDLE also expressed the highest level of beta-actin, a target cytoskeletal protein for S100A4. It is conceivable that the high expression of S100A4 in PDLE is a result of the maturation of the PDL and/or a response to mechanical stress generated by mastication. Since there was a marked difference of S100A4 expression between PDL and gingiva, we propose that S100A4 could be a useful marker for distinguishing cells from these two tissues.
In addition to seven known exons of the amelogenin gene, recent studies have identified two exons downstream of amelogenin exon 7 in genomic DNA of mouse and rat. Here the spatial and temporal expression of mRNAs and of the translated proteins derived from alternative splicing of the amelogenin gene ending with exon 8 and exon 9 were examined by in situ hybridization (ISH) and immunohistochemistry (IHC). RNA signals for exons 8 and 9 were expressed in the ameloblast layer extending from early presecretory to postsecretory transitional stages of amelogenesis. IHC of amelogenin proteins that include sequences encoded by these exons demonstrated identical localization of these proteins in the ameloblast layer corresponding to RNA signals identified by ISH. There was intense immunostaining of the enamel matrix secreted by these cells. Western blotting analysis of rat enamel proteins revealed three distinct protein bands with sequences encoded by the new exons. These data confirmed the existence of the transcripts of alternatively spliced mRNAs coding for exons 8 and 9 of the amelogenin gene in rat tooth germs and suggest that the translated proteins contribute to the heterogeneity of amelogenins and have some significant roles in enamel formation and mineralization.
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